By Kan Wang
A most suitable selection of Agrobacterium-mediated transformation concepts for cutting-edge plant genetic engineering, useful genomic research, and crop development. quantity 1 information the main up to date innovations on hand for twenty-six plant species drawn from cereal vegetation, commercial crops, legume crops, and vegetable vegetation, and offers a number of equipment for introducing DNA into 3 significant version plant species, Arabidopsis thaliana, Medicago truncatula, and Nicotiana. The authors additionally define the fundamental equipment in Agrobacterium manipulation and techniques for vector building. quantity 2 comprises one other thirty-three confirmed innovations for root vegetation, turf grasses, woody species, tropic vegetation, nuts and culmination, decorative vegetation, and medicinal vegetation. extra chapters supply tools for introducing DNA into non-plant species, reminiscent of micro organism, fungi, algae, and mammalian cells. The protocols persist with the winning tools in Molecular Biology™ sequence layout, each one delivering step by step laboratory directions, an advent outlining the foundations in the back of the procedure, lists of the required gear and reagents, and tips about troubleshooting and heading off recognized pitfalls.
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Extra resources for Agrobacterium Protocols, Second Edition: Volume I (Methods in Molecular Biology Vol 343)
And Lippincott, B. B. (1975) The genus Agrobacterium and plant tumorigenesis. Annu. Rev. Microbiol. 29, 377–405. 2. Allen, O. N. and Holding, A. J. (1974) Genus II. Agrobacterium, in Bergey’s Manual of Determinitive Bacteriology, 8th ed. (Buchanan, R. E. and Gibbons, N. ), Williams & Wilkins, Baltimore, pp. 264–267. 3. Kerr, A. (1992) The genus Agrobacterium, in The Prokaryotes, Vol. , Truper, H. ), SpringerVerlag, New York, pp. 2220–2235. 4. Snell, J. J. S. (1984) General introduction to maintenance methods, in Maintenance of Microorganisms: A Manual of Laboratory Methods (Kirsop, B.
When plasmid cointegration in A. tumefaciens is intended, triparental mating is much more efficient than electroporation, and these functions are necessary. When plants are transformed with various gene constructs, it is a good idea to always transform plants in parallel with such a vector, which is extremely useful in monitoring many aspects of transformation processes. Many factors are involved in successful transformation, and it is not a simple task to maintain capability of plant transformation stably over time.
Tumefaciens was described, revealed that derivatives of a relatively old vector, pBin19 (12), such as pBI121 (13), pIG121Hm (9), and others, were still used in about 40% of these studies. One reason could be that these vectors were widely circulated at early stages of plant transformation, and accumulated data in the literature from their use has built a lot of confidence. Another reason might be a convenient feature of pBI121, namely, that one-step replacement of the β-glucuronidase (Gus) gene with another gene can quickly create an overexpression vector for the gene.